Changchun University of Chinese Medicine, Changchun 130117, China.
Department of Urology, The Affiliated Hospital of Changchun University of Chinese Medicine, Changchun 130021, China.
Phytomedicine. 2023 Oct;119:155006. doi: 10.1016/j.phymed.2023.155006. Epub 2023 Aug 6.
Erectile dysfunction is common among the complications of diabetes mellitus. Shaofu Zhuyu decoction (SFZYD) is commonly used to treat diabetic mellitus erectile dysfunction (DMED). However, its main active components and specific mechanism are still unknown.
To confirm the activity of SFZYD in improving DMED, explore the main active components of SFZYD, and clarify the underlying mechanism.
A diabetic rat model was induced with streptozotocin (STZ). After intragastric administration, erectile function was assessed by the maximum intracavernous pressure (ICPmax)/mean arterial pressure (MAP). Corpus cavernosum fibrosis was evaluated by Masson staining, and ELISA methods were used to determine the serum levels of IL-6, TNF-α, IL-10, IL-4 and IL-1β to evaluate inflammation. Then, the main active components of SFZYD were identified by UPLC‒MS/MS. Finally, the target and biological mechanism of SFZYD in improving DMED were predicted by combined network pharmacology and transcriptomics, which was also validated by molecular docking and cellular thermal shift assay (CETSA) experiments.
SFZYD significantly improved erectile dysfunction and inhibited inflammatory responses and local tissue fibrosis in diabetic rats. A total of 1846 active components were identified by UPLC‒MS/MS, and isorhamnetin was the main active component. The transcriptomic results were used to identify differentially expressed genes among the control, DM and SFZYD groups, and 1264 differentially expressed genes were obtained from the intersection. The network pharmacology results showed that SFZYD acts on core targets such as AKT1, ALB, HSP90AA1 and ESR1 through core components such as isorhamnetin, quercetin and chrysophanic acid. Further combined analysis revealed that multiple targets, such as CYP1B1, DPP4, NOS2 and LCN2, as well as the regulation of the PI3K-AKT signaling pathway, may be important mechanisms by which SFZYD improves DMED. Molecular docking verification showed that isorhamnetin, the key component of SFZYD, has good binding ability with several core targets, and its binding ability with CYP1B1 was the strongest. The CETSA results showed that isorhamnetin binds to CYP1B1 in CCECs.
SFZYD improves DMED, inhibits the inflammatory response and alleviates local tissue fibrosis. The combined application of transcriptomic, network pharmacology, molecular docking and CETSA approaches was helpful for revealing the mechanism by which SFZYD improves DMED, which may be related to the regulation of CYP1B1 and the PI3K-Akt signaling pathway.
勃起功能障碍是糖尿病患者常见的并发症之一。少腹逐瘀汤（SFZYD）常用于治疗糖尿病性勃起功能障碍（DMED）。然而，其主要的活性成分和具体的作用机制尚不清楚。
确认 SFZYD 改善 DMED 的活性，探索 SFZYD 的主要活性成分，并阐明其潜在的作用机制。
采用链脲佐菌素（STZ）诱导糖尿病大鼠模型。通过最大海绵体内压（ICPmax）/平均动脉压（MAP）评估经胃给药后的勃起功能。通过 Masson 染色评估海绵体纤维化，酶联免疫吸附试验（ELISA）方法测定血清白细胞介素 6（IL-6）、肿瘤坏死因子-α（TNF-α）、白细胞介素 10（IL-10）、白细胞介素 4（IL-4）和白细胞介素 1β（IL-1β）水平，以评估炎症反应。然后，通过 UPLC-MS/MS 鉴定 SFZYD 的主要活性成分。最后，通过整合网络药理学和转录组学预测 SFZYD 改善 DMED 的靶标和生物学机制，并通过分子对接和细胞热转移分析（CETSA）实验进行验证。
SFZYD 显著改善了糖尿病大鼠的勃起功能障碍，并抑制了炎症反应和局部组织纤维化。通过 UPLC-MS/MS 鉴定出 1846 种活性成分，异鼠李素是主要的活性成分。转录组学结果用于鉴定对照组、DM 组和 SFZYD 组之间的差异表达基因，从交集获得 1264 个差异表达基因。网络药理学结果显示，SFZYD 通过异鼠李素、槲皮素和大黄酸等核心成分作用于 AKT1、ALB、HSP90AA1 和 ESR1 等核心靶点。进一步的联合分析表明，CYP1B1、DPP4、NOS2 和 LCN2 等多个靶标以及 PI3K-AKT 信号通路的调节可能是 SFZYD 改善 DMED 的重要机制。分子对接验证表明，SFZYD 的关键成分异鼠李素与几个核心靶点具有良好的结合能力，与 CYP1B1 的结合能力最强。CETSA 结果表明，异鼠李素在 CCECs 中与 CYP1B1 结合。
SFZYD 改善 DMED，抑制炎症反应，减轻局部组织纤维化。转录组学、网络药理学、分子对接和 CETSA 方法的联合应用有助于揭示 SFZYD 改善 DMED 的作用机制，可能与 CYP1B1 和 PI3K-Akt 信号通路的调节有关。
